Groves Jay T, Parthasarathy Raghuveer, Forstner Martin B
Department of Chemistry, University of California, Berkeley, CA 94720, USA.
Annu Rev Biomed Eng. 2008;10:311-38. doi: 10.1146/annurev.bioeng.10.061807.160431.
Imaging membrane dynamics is an important goal, motivated by the abundance of biochemical and biophysical events that are orchestrated at, or by, cellular membranes. The short length scales, fast timescales, and environmental requirements of membrane phenomena present challenges to imaging experiments. Several technical advances offer means to overcome these challenges, and we describe here three powerful techniques applicable to membrane imaging: total internal reflection fluorescence (TIRF) microscopy, fluorescence interference contrast (FLIC) microscopy, and fluorescence correlation spectroscopy (FCS). For each, we discuss the physics underpinning the approach, its practical implementation, and recent examples highlighting its achievements in exploring the membrane environment.
对膜动力学进行成像研究是一个重要目标，其背后的原因在于细胞膜上或由细胞膜所协调的大量生物化学和生物物理事件。膜现象的短长度尺度、快速时间尺度以及环境要求给成像实验带来了挑战。多项技术进展提供了克服这些挑战的方法，在此我们描述三种适用于膜成像的强大技术：全内反射荧光（TIRF）显微镜、荧光干涉对比（FLIC）显微镜以及荧光相关光谱（FCS）。对于每种技术，我们将讨论其背后的物理原理、实际操作方法，以及近期展示其在探索膜环境方面所取得成果的实例。
